The present invention relates to an adsorbent for removing from body fluid at least one interleukin (hereinafter referred to as "IL") selected from the group consisting of interleukin-1 (hereinafter referred to as "IL-1"), interleukin-2 (hereinafter referred to as "IL-2"), interleukin-6 (hereinafter referred to as "IL-6") and interleukin-8 (hereinafter referred to as "IL-8"); a method for removing the above-mentioned IL(s) in body fluid by means of the above-mentioned adsorbent; an adsorber for removing the above-mentioned IL(s) by means of the above-mentioned adsorbent; an adsorbent for removing tumor necrosis factor (hereinafter referred to as "TNF"); a method for removing TNF from body fluid by means of the above-mentioned adsorbent; and an adsorber for removing TNF by means of the above-mentioned adsorbent.
An immunocompetent cell produces various kinds of active substances when causing immune response. One portion thereof is a proteinous substance called a cytokine and plays a greatly important role as a biophylactic factor which is closely related to various kinds of antigen-specific response and/or non-specific inflammatory response. Essentially, a cytokine is necessary and indispensable for maintaining biological homeostasis and is produced excessively in pathological conditions such as inflammation and the like, relating to the formation and the prolongation of inflammation and the like.
IL-1, the gene thereof was cloned in from 1984 to 1985, is a proteinic factor having a molecular weight of about 17 kD which is produced mainly from cells of the monocyte/macrophage lineage. As IL-1, there exist IL-1.alpha. and IL-1.beta. which are originated from different genes, respectively. The activated macrophage produces IL-1 .alpha. and IL-1.beta. in a IL-1.alpha.: IL-1.beta. ratio of about 1: 9.
It has been made clear that IL-1 plays an important role in all kinds of biological reactions such as immune, inflammation, hematopoiesis, secretion in the nerve, biological homeostasis and the like. Contrary to this, it has been shown that an abnormal production of IL-1 causes various kinds of diseases. For example, there is autoimmune disease as one of the diseases and it has been shown that IL-1 relates to the formation of inflammation of connective tissue diseases causing a systemic chronic inflammation and, among them, particularly to the formation of inflammation of rheumatoid arthritis (hereinafter referred to as "RA"). IL-1 has a cartilage destroying function by causing overproduction of prostaglandins and collagenase from synovial cells and chondrocytes, and a bone resorbing function by causing activation of osteoclasts, and it has been strongly shown that there is a possibility that IL-1 relates to the formation of rheumatoid joints. Then, it has been reported that when IL-1 is injected into a cavitus articulare of a treated animal, fugitive arthrisis can be reappeared. And, it has been shown that IL-1 plays a leading role in the pathogenesis of RA. Further, in recent years, the followings have been reported; in diseases which are included by the concept such as systemic inflammatory response syndrome (hereinafter referred to as "SIRS"), inflammatory cytokines such as IL-1 and the like are produced excessively, and the systemic inflammatory response proceeds mainly because of functions of these cytokines; then, tissular disorders and failures of many organs occur and, sometimes, a death is caused. Further, a higher concentration of IL-1 has been detected in an inflammatory site or in peripheral blood of a patient with lupus erythematosus, Lyme disease, osteoporosis, Kawasaki disease, gouty arthritis, endometritis or premature labor than that of a normal human, and it has been shown that IL-1 relates closely to the formation of the above-mentioned inflammation in these diseases. Further, it has been shown that IL-1 is produced in a patient on dialysis because of various kinds of factors, and that IL-1 relates closely to the pathogenesis of dialytic complications including dialysis related amyloidosis. Also, IL-1 has a function for accelerating the production of other cytokines in addition to the above-mentioned functions, and it is confirmed that IL-1 is a main causative substance of vicious circle in inflammation. Though IL-1 relates closely to the inflammation of each kind of diseases, the present situation is that any effective method of inhibiting the functions of IL-1 or of removing IL-1 from body fluid of patients with the above-mentioned diseases has not been established.
Also, IL-2 is an active factor found by Morgan et al in 1976 as T cell growth factor (TCGF) capable of maintaining T cell for a long term, from an activated culture medium obtained by culturing lymphocytes of peripheral blood with an antigen, mitogen or the like. It has been gradually made clear in later researches that this active factor has an activity to accelerate the division of a thymocyte, to activate a cytotoxic T cell, to derive the differentiation of a B cell, to activate a natural killer (NK) cell, and to derive an activity of a lymphokine activated killer (LAK). The TCGF was named IL-2 uniformly in 1979. And then, the gene of IL-2 was cloned by Taniguchi et al in 1983 and the primary structure thereof was made clear.
IL-2 is produced mainly by a T cell, acts on cells with IL-2 receptor (IL-2R) on the surface thereof, such as a T cell, a B cell, a NK cell, a monocyte, a macrophage, a glioma cell and the like, and has various functions to cause proliferation, differentiation, activation and the like of the above-mentioned cells. It has been shown, however, that an abnormal production of IL-2 has a harmful function on a living body. For example, it is known that a cytokine exists in blood of a patient with sepsis in an abnormally high concentration. When sepsis becomes serious, the so-called "septic shock" occurs. This septic shock can be classified to two types. As one type, it has been reported that IL-2 exists in an abnormally high concentration and relates to the formation of inflammation thereof (refer to S. Endo et al, Circulatory Shock, 38, pages 264-274 (1992)). Also, it has been reported that among septic shocks, the septic shock to which IL-2 relates has a bad prognosis. As described in the above, though IL-2 relates closely to the formation of septic shock, the present situation is that any method of inhibiting the functions of IL-2 or of removing IL-2 from body fluid has not been established.
Further, IL-6 was isolated and purified by Kishimoto et al in 1985 as a factor to derive only the production of antibody without causing the acceleration of proliferation of an activated B cell. And, IL-6 is a cytokine of which cDNA was isolated and of which whole base sequence was determined by Hirano et al in 1986.
IL-6 has many biological activities, for example, to cause the derivation of the production of acute phase protein in an immunocompetent cell and a hepatic cell.
IL-6 is produced from each kind of various cells such as a monocyte, a fibroblast, an angioendothelial cell and a skin keratinocyte when the stimulation of various kinds of inflammatory substances including lipopolysaccharide (LPS) are added thereto. And, therefore, chronic inflammatory response occurs by an overproduction of IL-6 because IL-6 has a function for enhancing the inflammatory response. For example, it has been reported that IL-6 is produced by a B lymphoblast in centroblast of enlarged lymph nodes of Castleman disease, and the improvement of clinical symptom and the decrease of serum level of IL-6 are caused by a surgical removal of the lesional lymph nodes. In further recent years, it has been reported that in diseases included by the above-mentioned concept SIRS, the concentration of inflammatory cytokines such as IL-6 in blood is high, and systemic inflammatory response proceeds mainly because of these functions similarly in the case of IL-1, then tissular disorders and failures of many organs occur and, sometimes, a death is caused. An abnormally higher concentration of IL-6 is detected in an inflammatory site or in peripheral blood of a patient with autoimmune diseases such as RA, systemic lupus erythematosus, chronic diseases with proliferation such as mesangial nephritis and psoriasis and, further, dialytic complications such as dialysis related amyloidosis than in that of a normal human. And, it is considered that IL-6 relates closely to the formation of inflammation of those diseases. It is, however, the present situation is that any effective method of inhibiting the functions of IL-6 in body fluid or of removing IL-6 from body fluid has not been established.
IL-8 is a cytokine purified as monocyte-derived neutrophil chemotactic factor (MDNCF), and gene thereof was also cloned by Matsushima et al in 1987. According to later researches, IL-8 has chemotaxis not only to a neutrophil but also to a basophil and at T lymphocyte. IL-8 is produced by various kinds of cells such as a monocyte, a macrophage, a fibroblast, an angioendothelial cell, a chondrocyte and the like.
The infiltration of a neutrophil and a lymphocyte can be reappeared even in vivo by intracutaneous/subcutaneous and intra-articular administration of IL-8.
To maintain the administration of a large amount of IL-8 is remarkably harm to tissues, and causes the destruction of tissues of adult respiratory distress syndrome in an alveolus and the destruction with the infiltration of a large amount of lymphocytes in an arthrosis. Experimentally, IL-8 relates essentially to the infiltration of a neutrophil in dermatitis derived by lipopolysaccharide and during reperfusion after ischemia. And, it has been proved that the destruction of tissues can be almost completely inhibited by a neutralizing antibody against IL-8. Further, an abnormally higher concentration of IL-8 has been detected in an inflammatory site or in peripheral blood of a patient with diseases such as RA, gouty arthritis, psoriasis, contact dermatitis, idiopathic fibroid lung, adult respiratory distress syndrome, inflammatory bowel disease, immune angiitis, glomerular nephritis, urinary tract infection, cardiac infarction, asthma, respiratory tract infection, perinatal infectious disease, rejection in organ transplantation and the like, than in that of a normal human (refer to Menekiyakuri, 12, No. 1, pages 15-21 (1994)). In further recent years, it has been reported that in diseases included by the above-mentioned concept SIRS, the concentration of inflammatory cytokines such as IL-8 in blood is high, and systemic inflammatory response proceeds mainly because of those functions similarly to in the cases of IL-1 and IL-6, and then tissular disorders and failures of many organs occur and, sometimes, a death is caused. Further, IL-8 is produced in a patient on dialysis because of various kinds of factors, and it has been shown that IL-8 relates closely to the pathogenesis of dialytic complications including dialysis related amyloidosis. It is, however, the present situation is that any effective method of inhibiting the functions of IL-8 in body fluid or of removing IL-8 from body fluid has not been established.
Then, TNF can be mainly classified into a tumor necrosis factor originated from cells of the monocyte/macrophage lineage (hereinafter referred to as "TNF.alpha.") and into a tumor necrosis factor originated from a lymphocyte (hereinafter referred to as "TNF.beta."). It was reported by Carswell et al in 1975 that TNF.alpha. was found as a biologically active substance which appears in blood when Bcillus Calmette-Guerin (BCG) was administered to a CD-1 Swiss mouse and, then, after two weeks a bacterial mitogen was administered. The amino acid sequence thereof was made clear by Aggarwa et al. Also, the amino acid sequence and gene arrangement of human TNF.alpha. were made clear by Pennica et al, Shirai et al and Wang et al in 1985. TNF.beta. was reported by Granger et al in 1968 as a factor which gives damage not to a normal cell but only to a tumor cell or as a factor which inhibits the growth of a tumor cell. And, human-type cDNA of TNF.beta. was cloned and the structure thereof was determined by Gray et al in 1984. Though TNF.beta. was also called lymphotoxin (LT), TNF.beta. has been called TNF.beta. because TNF.beta. has about 30% of homology to TNF.alpha.. Though both of TNF.alpha. and TNF.beta. bind to the same receptor and it is considered that they have an approximately same activity, TNF.alpha. has a higher activity and a slightly different activity has been found between them recently.
According to the recent researches, it has been made clear that the effect of TNF relates not only to antineoplastic activity but also to immune, inflammation, fat metabolism, coagulating and fibrinolysis cascades, hemopoiesis and the like. From the researches until now an abnormally higher concentration of TNF is detected in an inflammatory site or in peripheral blood of a patient with a disease such as RA or arterioslerosis than in that of a normal human. And the relation between TNF and these diseases has been shown. In further recent years, in diseases included by the above-mentioned concept SIRS, an inflammatory cytokine such as TNF or the like is produced excessively, a systemic inflammatory response proceeds mainly because of these functions, and then tissular disorders and failure of many organs occur and, sometimes, a death is caused. Further, TNF is produced in a patient on dialysis because of various kinds of factors, and it has been shown that TNF relates closely to the pathogenesis of dialytic complications including dialysis related amyloidosis. Also, in addition to the above-mentioned functions, TNF has a function for accelerating the production of other cytokines and is considered to be a main causative substance of the vicious circle of an inflammatory site. It is, however, the present situation is that any effective method of inhibiting the effect of TNF in body fluid or of removing TNF from body fluid has not been established.
In order to efficiently adsorb at least one IL selected from the group consisting of IL-1, IL-2, IL-6 and IL-8 which are present in body fluid and efficiently adsorb TNF which is present in body fluid, an adsorbent which can adsorb the above-mentioned IL(s) and an adsorbent which can adsorb TNF were studied. As a result, it was found that a material which comprises a porous water-insoluble carrier and a compound satisfying the value of log P of at least 2.50, in which P is a distribution coefficient in an octanol-water system, and being immobilized onto the carrier, can efficiently adsorb the above-mentioned IL(s) and TNF in body fluid. Then, the present invention has been accomplished.
An object of the invention is to provide an adsorbent which can efficiently adsorb at least one IL selected from the group consisting of IL-1, IL-2, IL-6 and IL-8 which are present in body fluid.
A further object of the invention is to provide a process for removing the above-mentioned IL(s) in body fluid by means of the above-mentioned adsorbent.
A still further object of the invention is to provide an adsorber for removing the above-mentioned IL(s) by means of the above-mentioned adsorbent.
Another object of the invention is to provide an adsorbent which can efficiently adsorb TNF in body fluid.
A further another object of the invention is to provide a method for removing TNF in body fluid by means of the above-mentioned adsorbent.
A still further another object of the invention is to provide an adsorber for removing TNF by means of the above-mentioned adsorbent.
These and the other objects of the present invention will become apparent from the description hereinafter.